Datasheet Extract

Application

Conjugation.

Background

The Link-A-Light HRP conjugation kit allows conjugations to be set up in seconds, simply by adding a solution of the protein to be labeled to the lyophilised HRP mixture (Figure 1).By circumventing the desalting or dialysis steps that commonly interrupt traditional protein conjugation procedures, Link-A-Light technology can be used to label small quantities of protein (see Principle).

Protocols

1. Before you add antibody to the Link-A-Light Mix, add 1 µl of Link-A-Light Modifier reagent for each 10 µl of Antibody to be labeled. Mix gently.2. Remove the screw cap from the vial of Link-A-Light Mix and pipette the antibody sample (with added Link-A-Light Modifier) directly onto the lyophilised material. Resuspend gently by withdrawing and re-dispensing the liquid once or twice using a pipette.3. Place the cap back on the vial and leave the vial standing for 3 hours at room temperature (20-25°C). Alternatively, and sometimes more conveniently, conjugations can be set up and left overnight, as the longer incubation time has no negative effect on the conjugate.4. After incubating for 3 hours (or more), add 1 µl of Link-A-Light Quencher FD reagent for every 10 µl of antibody used. The conjugate can be used after 30 minutes.

Storage

Storage of Conjugates: For any new conjugate, initial storage at 4°C is recommended. A preservative may be desirable for Long-term storage. Other storage conditions (e.g. Frozen at -70°C or stored at -20°C with 50% Glycerol) may also be satisfactory. The best conditions for any particular conjugate must be determined by experimentation.

Storage of Kit: The kit is shipped at ambient temperature in a tamper-evident polypropylene container. Store at -20°C upon receipt. Shelf life: one year from despatch.

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